7-substituted thio steroids



3,074,932 7-SUBSTITUTED THIO STEROID S Robert E. Schaub, Paramus, and Martin J. Weiss,

N.J., assignors to American Cyanamid Company, York, N.Y., a corporation of Maine No Drawing. Filed Aug. 15, 1960, Ser. No. 49,406 12 Claims. (Cl. 260-2395) Oradell, New

This invention relates to new steroid compounds. More particularly, the invention relates androstanes and androstenes.

The compounds of the present invention can be illustrated by the following structural formula:

consisting of lower alkylthio, lower alkylsulfinyl and lower alkylsulfonyl radicals, C

is a divalent radical of the group consisting of:

HO-CH 1 911(1 NH o: o: N-"O t radicals, I

Cl C4 is a. trivalent radical of the group consisting of: A] ,3.

I \CH/ 0112 and radicals' and when R is the methyl radical, R is a lower alkylthio radical,

is the I radical and when is the to 7-substituted thioa 3,074,932 Patented Jan. 22, 1963 alkyl mercaptan or a lower alkyl thiolicacid. These reactions can be specifically illustrated as follows:

--so-orr,

i komsn l v uoA -Hon g 1 Y OHi one o sou,

i more, usually three to four days.

" from about 3 to be illustrated specifically The reaction with lower alkyl thiolic acids is carried out by treating a solution of the A i-3-keto steroid with an excess of thiolic acid at a temperature within the range of from C. to C. for aperiodjof'time ranging about 24 hours. I

By either procedure the first step in isolation of prodnet is removal of solvent under reduced pressure. Usually the crude product thus obtained is in the form of a guru. Trituration with 'ethylether generally induces crystallization. The product can then be recrystallized from appropriate organic solvents. A combination of acetone and petroleum ether (GO-70 C. boiling range) is often useful for recrystallization. Crude gums resistant to crystallization may have to be subjected to preliminary chromatographic purification.

The :A -3-keto steroid starting materials are generally known compounds and are indicated by literature references in the examples hereinafter. The other starting materials are prepared by methods described in the chemical literature. These procedures involve either the treatment of a A -3-keto steroid with chloranil or the bromination of a A -3-keto steroid with N-bromsuccinimide to give a 6-bromo-A -3-keto steroid followed by dehydrobromination on treatment with hot collidine. These reactions can as follows:

g OC'CH,OH; O CHCH: H [j (\o I NBS I Br (IV) (V) leollidine O-fi-CHaCH: 0-fi-CH2CHs I 0 0 CHaSH HOA'cHCl \SOH: 01 01 (VII) (VI) and The lower alkylsulfinyl (sulfoxide) derivatives or this ()3 0H invention are prepared by treatin L L--OH, thio derivative with a slight excess over 0 HO OH alent'of monoperphthal-ic acid. This re F it 'out in an inert solvent, such as methylene chl temperature for about 24 hours. cm r-nfl (sulfone) derivatives are obtaine of the alkylsulfinyl derivatives w The latter oxidation usually requires about 48 hours of reaction time. The following (VIII) reactions:

40 on "CH HOl Monoperphthalie F x ac'd 0- (MILA) 0 (MFA) N 0:

OHa

wherein Ac is as defined above.

The novel androstane derivatives 0 f the present invention can be prepared from 4-androstene-3 -ketones such 3-one which is ostane steroid g as17-acetylo1ry-7otethylthio-4-an'drostene- Br reacted with lithium in liquid ammonia to sa :(XI) (XII) double bond and thereby produce an andr having the trans A/B ring junct leomdme illustrated as follows: OH OH 0A0 OH, I CH: W11] .0 1 I Li, 1. NH:

I +cHisH I or i 0 o (X'Iv) mm txvm) us. 2,908,694)

(XIX) defined above. Introduction of the hydroxyrnethylene wherein Ac is as group at -2 is treating the 4,5-dihydro-3-ketone with an alkyl formate, for example ethyl formate, in the presence of a strong base, such as sodium hydride or sodium alkoxide in benzene at room temperature for a prolonged eriod such as, for example, five days. the hydroxymethylene derivative with hydrazine hydrate to give the pyrazole derivative is easily carried out and can be effected by brief treatment, fifteen minutes usually being sufiicient in ethanolic solution at about room temperature. The products of this reaction are obtained by filtration or evaporation of solvents and are purified by crystallization and/ or recrystallization techniques well carried out by known in the art.

The hydroxyrnethylene derivatives and the pyrazolo derivatives of this invention may exist in several tautorneric forms. Thus, the product designated as the hydroxymethylene derivative may exist in any one or all of the following structures: Y

The pyrazolo derivative may have any of the following tautomeric forms (D, E and F).

a 'H-N N (D) The use of structure D in the specification and claims is intended to include tautomeric forms.

The process of this invention proceeds under steric influences and steroisomers are formed. However, one isomer is obtained in predominant amount. The predominant isomer has in each case been characterized herein as possessing the alpha configuration for the 7-lower alkylthio, lower alkanoylthio, lower alkylsulfinyl or lower alkylsulfonyl group. This configuration has been designated in order to provide a more complete exposition of the present invention and in order that the specification shall constitute a more useful contribution to the art. However, the designated configuration of the 7-lower alkylthio, lower alkanoylthio, lower alkylsulfinyl or lower alkylsulfonyl group is based upon an analysis of molecular rotation data presently appearing in the chemical literature and is therefore not to be interpreted except in relation to the state of the art presently known to organic chemists. It will be apparent that no part of the specifica- .tion will be materially defective if it should later be established that the configuration is the opposite of that deducible from data presently available to workers in the field.

ticularly useful as androgenic and anabolic agents. may be used reaction mixture ,ml. fraction contains the desired product. is evaporated '6 The new compounds of the present invention are par- They inandrogenic-replacement therapy, in supportive therapy for patients suffering from debilitating diseases and in the treatment of post-operative trauma.

The following examples illustrate in greater detail the preparation of the 7 -substituted thioandrostanes and thioandrostenes of the present invention.

EXAMPLE I Preparation of 7u-Acetylthi0-19-Nor-Testosterone Acetare (1 7B-Acet0xy-7a-Acetylthi0-19-N0r-4-Androsten- 3-0ne, II)

A solution of 500 mg. of 17(3-acetoxy-19-nor-4,6-androstadien-S-one (I) [L. Velluz, B. Goffinet, and G. Arniard, Tetrahedron, 4, 241 (1958)] in 2 ml. of freshly distilled thiolacetic acid is heated under reflux for two hours. The is evaporated to dryness under reduced pressure. The residual glass is dissolved in 15 cc. of benzene and chromatographed on 50 g. of silica gel. The column is washed with 300 ml. of 2% ether-in-benzene, 300 ml. of 4% ether-in-benzene, 300 ml. of 6% ether-inbenzene, 350 ml. of 8% ether-in-benzene; these washings are discarded. The column is then eluted with 500 ml. of 10% ether-in-benzene', the eluate is evaporated to dryness under reduced pressure to give 200 mg. of a glass.

The glass is dissolved in 5 ml. of the lower and 5 ml. of the upper phases of the system heptane: methyl Cellosolve and the liquid is mixed with 10 g. of diatomaceous earth. This mixture is packed on top of a column which has been prepared from 100 g. of diatomaceous earth and 50 ml. of the lower phase of the solvent system just described. The column is eluted with the upper phase and the effluent is allowed to pass through a recording spectrophotometer which has been set at 238 mg. The first 950 ml. of effiuent contains little ultraviolet-absorbing material and is discarded. The next 330 ml. contains much ultraviolet-absorbing material. This fraction is taken to dryness to give 136 mg. of product as an amorphous solid;

[a] -59(1.O3% in ouch); xsgzg 238 my (e=l8,300); xiii; 5.71, 5.87, 6.12, 8.00 1.

EXAMPLE II Preparation of 7or-Methylthi0-19-N0r-Test0sterone Acetate (17B-Acetoxy-h-Methylthio-I9-Nor-4-Androsten- 3-One, III) A solution of 500 mg. of l7fi-acetoxy-19-nor-4,6-andro stadien-3-one (I), 1 ml. of concentrated hydrochloric acid and 5 ml. of methyl mercaptan in 35 ml. of glacial acetic acid is allowed to stand at 5-8 C. for four days. The reaction mixture is evaporated to dryness under reduced pressure, and the residue is taken up with methylene chloride and washed with'excess saturated sodium bicarbonate solution and then with water. The organic phase, dried with anhydrous magnesium sulfate is evaporated to dryness under reduced pressure to give 400 mg. of a glass. This glass is subjected to partition chromatography on diatomaceous earth by the procedure described above in Example I. The system petroleum ether (31. -100 C.) methanol-2-methoxyethanol (821:1) is used. The column is packed with 300 g. of diatomaceous earth and the recording spectrophotometer is set at 240 mg. The first 950 ml. of effluent contains only small amounts of ultraviolet-absorbing material and is discarded. The next 400 This fraction to dryness under reduced pressure to give a glass, crystallization of which from acetone-petroleum ether (20-40") gives 90 mg. of white crystals, melting point 123-125" C. Two recrystallizations from the same 21111. of a 10% 'zene, is washed successively with 8% 7 EXAMPLE III Preparation of 4-ChZora-7a-Methylthi0test0sterone Prpionate (4-Chl0r0-7a-Methylthi0-]7p Proprionoxy 4- Andr0sten-3-One, VII) A solution of 4-chlorotestosterone propionate (IV) N. Kirk, and V. Petrow, Brit. Patent 120 ml. of reagent carbon tetrachloride is concentrated by distillation to about 100 ml. to remove moisture. To the cooled solution is added solution of reagent pyridine in carbon tetrachloride. After flushing with carbon dioxide (bone dry) 1.03 g. of -bromosuccinimide is added and the solution is heated under reflux for 80 minutes while [a] 185(1.1% in oHOn); its? 266 mu (611,200); A 2.95, 3.41, 5.80, 5.93, 7.9

max.

6-bromo-4-chlorotestosterone propionate (V) (1.5 g.) suspended in 15 ml. of syrn. collidine and refluxed under for 30 minutes, solution at the boiling point. The cooled solufrom collidine hydrobromide and the mother liquid, after being diluted with 100 ml. of ber1- ated aqueous sodium bicarbonate and Water, dried with pressure and the crude residual solid is crystallized from petroleum ether to yield 546 mg. (44%) of 6-dehydro-4- chlorotestosterone propionate (VI), melting point 100- 102. Recrystallization from the same solvent gives white crystals, melting point 103-104;

[a]n +79(0.4l% in CHCIs); M32 295 mp. (624,600) k 5.73, 5.96, 6.20, 6.42, 8.411.

with water, dried with anhydrous magnezium sulfate and evaporated to dryness. Crystallization of the residual syrup from acetone-petroleum ether gives 85 mg. of product (VII), melting at 235-240 C. Recrystallization from the same solvents gives white crystals, melting point 252-254" C. (dec.);

A3 52 255 m (613,800) A2,; 5.72, 5.90, 6.28, 840

EXAMPLE IV Preparation of 7a-Acetyltlzi0-9a-Flu0r0-11;8-Hydroxy- 17a-Methyltest0ster0ne 7 a-A cetyllhi0-9u-Flu0r0-1 J p,

17p-Dihydr0xy-I7a-Methyl-4-Arzdr0sten-3-One, X)

To a solution of Qa-FIHOIO-l1,8,17fidihydrOXy-17amethyl-4-androsten-3-one (VIII) (1.0 g.) [M. E. Herr, et al., J. Am. Chem. Soc. 78, 501 (1956)] in 100 ml. of dry fibutanol, freed from moisture by distillation of 30 ml. of solvent is added -1 g. of chloranil (2,3,5,6-tetra- 'terone (XIII), melting chlorobenzoquinone) and 10 mg. of p-toluenesulfonic acid. The mixture is refluxed, under an nitrogen for 20 hours, solution being complete in approximately 45 minutes. The solution is evaporated to dryness under reduced pressure. The residual syrup is dissolved in methylene chloride and the solution is Washed with three portions of ice-cold 5% sodium hydroxide solution and then with water (to neutrality), dried with and evaporated to dryness under reduced pressure leaving an amorphous solid. Crystallization from acetone-petroleum ether gives 200 mg. of 6-dehydro-9u-fluoro-11B hydroxy 17oz methyltestosterone (IX) melting point 239 C. dec. Recrystallization from the same solvents gives white crystals, melting point 242 C. dec.; [u] +53.6(0.6% in MeOH); Mfg-2 A533,, 2.79, 2.88, 6.03, 6.18, 6.31

A suspension of 6-dehydro 9u-fiuoro-l lfi-hydroxy-lhmg.) in 1 ml. of thiolacetic approximately 25 minutes. The solution is evaporated to dryness under reduced pressure. Trituration of the residual solid with ether gives 88 mg. (72%) of product (X), melting point 241 C. dec. Several recrystallizations from acetone-petroleum ether gives white crystals melting point 243-246 C. dec.;

[oz] 4-1(0.24% in CHCl AEEQ A 2.91, 5.94, 6.04, 6.15, 8.0,0.

EXAMPLE V Preparation of 7 oz M ethylthz'o 17a Melhyltestosterone (17,8 Hydroxy 17oz Methyl 7a Methylllzio 4- Andr0sten-3-One, XIV) ml. to remove moisture. oxide, 1.77 g. of N-bromosuccimmide is added and the solution is refluxed while watt frosted bulb and under an atmosphere of carbon dioxide, for 40 minutes. The solution shows a negative test with moist potassium iodide-starch test paper. The hot solution is filtered and the filtrate is concentrated to dryness under reduced pressure. The residue is taken up in methylene chloride and washed with water, dried lization from acetone-petroleum ether 70) furnishes 2 g. (53%) of 6-bromo-17u-methyltestosterone (XII), melting point l32 recrystallizations from crystals, melting point 139-140 C. dec.;

[a] 26(0.8% in CHCl AEQQ 246 my (612,000); A 2.88, 5.93, 6.20

A solution of 1.5 g. of 6-bromo-17a-methyltestosterone .(XII) in 20 ml. of sym-collidine is refluxed, under an is washed (v./v.) sulfuric acid, saturated aqueous sodium bicarbonate and water, dried with anhydrous magnesium sulfate and evaporated to dryness under reduced pressure leaving a semi-solid. Recrystallization from acetone-petroleum ether (boiling point 6070 C.) furnishes 700 mg. (59%) of 6-dehydro-l7u-methyltestospoint C. (gas). Two more recrystallizations from the same solvent pair gives white crystals, melting point 194-195 C.;

[a] +33 (0.5% in CHCI3); A9552 283 mu (e=27,200)g REL; 2.91, 6.06, 6.21, 6.34;;

Ame 241 mp (614,600); REE, 2.83, 6.03, 6.19 1. EXAMPLE VI Preparation of 17,8-Acetoxy-7a-Methylthioandrostan- 3-One (XVIII) A solution of 2.0 g. of 17fi-acetoxy-7a-methylthio-4- androsten-S-one (XV) [R. E. Schaub and M. J. Weiss US. Patent 2,908,694 (1959)] in 25 ml. of dioxane and 25 ml. of ether is added dropwise to a solution of 300 mg. of lithium in 250 ml. of liquid ammonia over a period of 15 minutes. After stirring for an additional 35 minutes the blue color of the solution is discharged by the addition of 6.0 g. of ammonium chloride and the ammonia is allowed to evaporate. The residue is distributed between ether and water and the water extract is washed several times with methylene chloride. The combined organic extracts are washed with water, dried with anhydrous magnesium sulfate and evaporated to dryness under reduced pressure leaving 1.6 g. of amorphous material. Since at least partial deacetylation occurs (l7fi-hydroxy-7a-methylthioandrostan-3-one is present) as indicated by hydroxyl absorption at 289 in the infrared, the material'is acetylated with acetic anhydride in pyridine solution at room temperature overnight. After dilution with water, the mixture is extracted with methylene chloride, washed with water, dried with anhydrous magnesium sulfate and evaporated to dryness under re duced pressure leaving a syrup. Trituration with ether and filtration gives 720 mg. (36%) of 17fl-acetoxy-7amethylthioandrostan-3-one (XVIII) as a white powder, melting point 175-179 C. The material has no ultraviolet absorption;

[a] -44.4: (0.61% in CHCIa); REE; 5.75, 5.80, 8.00a EXAMPLE VII Preparation of 17fi-Hydroxy-2-Hydr0xymethylane-7a Methylthioandrostan-S-One (XIX) Sodium hydride (2.24 g. of a dispersion in oil) is added to a solution of 2.8 g. of 17/3-acetoxy-7a-methylthioandrostan-3-one (XVIII) (Example VI) in 100 ml. of reagent benzene and 5.6 ml. of ethyl formate freed from moisture by azeotropic distillation of 20 ml. The reaction mixture is allowed to stand under nitrogen for five days. Methanol (5.6 ml.) is added to decompose the excess hydride and the solution is then diluted with 280 ml. of benzene and 280 ml. of water. The layers are separated and the aqueous phase is extracted with benzene. The aqueous layer is then acidified with dilute hydrochloric acid and the precipitated enol is extracted with ether. The extracts are washed with water, dried with anhydrous magnesium sulfate and evaporated to dryness under reduced pressure to give a glass. Recrystallization from acetone-petroleum ether gives 1.6 g. (60%) of crystalline product (XIX), melting point 130- 140 C. (gas). The compound gives a deep red color with 1% alcoholic ferric chloride solution. Recrystalization from ethyl acetate gives white crystals, melting point 163-165 C. (previous softening);

[a] 29.2(1.4% in (II-I013); H52 282 III/.L (c7300); A5,; 2.88, 6.10, 6.2511.

EXAMPLE VIII Preparation of 17fi-Hydroxy-7a-Methylthioandrostano [3,2-c1Pyraz0le (XX) To a solution of 200 mg. of 17fi-hydroxy-2-hydroxymethylene-7a-methylthioandrostan-S-one (XIX) (prepared in Example VII) in 7 ml. of absolute alcohol is added 3.0 ml. of absolute alcohol containing 0.03 ml. of 98-100% hydrazine hydrate. The solution becomes turbid and a crystalline material separates after 10 minutes. The mixture remains colorless when tested with 1% alcoholic ferric chloride solution. The mixture is cooled and filtered to give 181 mg. of product (XX) in two crops, melting Several recrystallizations from acetone-water give white crystals, melting point 174 C. (gas);

[uln -22.2 (0.4% in 011013); m2, 233 mn (64750); REE}, 3.05, 6.90, 1046a EXAMPLE IX Pr paration of 7a-Methylsulfinyltestosterone 17-Acetate (XVI) To a solution of 7a-methylthiotestosterone 17-acetate (XV) (US. Patent 2,908,694) (500 mg.) in 10 ml. of methylene chloride is added 1.1 molar equivalents of monoperphthalic acid. The reaction mixture, protected from moisture, is allowed to stand at room temperature for 24 hours, during which period phthalic acid separates. The solution shows a negative test with 20% aqueous potassium iodide solution. The phthalic acid is collected by filtration. The filtrate is washed with dilute aqueous sodium carbonate solution and then with water. After drying over anhydrous sulfate, the solvent is evaporated at reduced pressure. The residue is triturated with petroleum ether (boiling point -70) and the product (XVI) (77%) 'is collected by filtration. For analysis, the product is recrystallized from acetonepetroleum ether, melting point ISO-152 C.;

[a]D-6.8 (1% in chloroform); A252 246 mp. (611,000); REE; 5.75, 5.97, 6.15, 8.00, 9.04;;

EXAMPLE x Preparation of 17i3-Acet0xy-7a-Methylsulfinylandrostan- This product is obtained in 66% yield by the procedure of Example IX from 17a-acetoxy-7a-methylthioandrostan-3-one (Example VI), The product is recrystallized from acetone-petroleum ether (boiling point 60-70); melting point 205-207 CL;

a snm (1%111 chloroform); x52; 9.6011 I i EXAMPLE XI Preparation of 7a-Methylsulfonyltestosterone 17-Acetate (XVII) This product is obtained in yield from la-methylsulfinyltestosterone 17-acetate (XVI) (Example IX) by the procedure of Example IX. The reaction time is 48 hours. The product is recrystallized from acetone petroleum ether (boiling point 60-70); melting point 165168 C.;

[a]n+19.7 (1.1% in chloroform); AXE? 241 my. (15,100); x55; 5.77, 6.00, 6.16, 7.33, 8.05, 8.88, 13.051.

EXAMPLE XII Preparation of 17,6-Acetoxy-7a-Methylsalfonylandrostan- 3-One [a]1 -4I.5 (0.9% in chloroform); A511, 5.75, 5.84, 7.73,

We claim:

1. The compound 4chloro-7a-methylthiotestosterone propionate.

2. The compound 17 3-hydroxy-2-hydroxymethylene- 7a-methylthio-androstan-3-one.

3. The compound 17,8-hydroxy-7a-methylthioandrostano-[3,2-c]pyrazole.

l7rx-methyltestosterone.

5. The compound 7a-methylsulfinyltestosterone-17- acetate.

6. The compound 7a-methylsulfonyltestosterone l7- acetate.

7. Compounds of the formula:

in which R is lower alkanol and R is lower alkyl-sulfinyl.

8. Compounds of the formula:

in which R is lower alkanol and R is lower alkyl-sulfinyl.

9. Compounds of the formula:

ORs

in which R is lower alkanol and R is lower alkyI-sulfonyl.

10. Compounds of the formula:

R 5 /\l l 10 0= "R0 in which R is lower alaknol and R is lower alkyl-sulfonyl.

References Cited in the file of this patent UNITED STATES PATENTS 2,908,694 Schaub etal Oct. 13, 1959 OTHER REFERENCES Dodson et al.: Journ. 'AmrChem. 800., vol. 81, page 1227, Mar. 5, 1959.

I UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. 3,07%932 January 22, 1963 Robert E, Schaub et al.

are in the above numbered patd that error appe Letters Patent should read as It is hereby certifie at the said ent requiring correction and th corrected below.

Column 3, lines 65 to 75 formula XIV should appear. as

shown below instead d f as in the patent:

column 11, lines 15 to 23 the formula should appear .as shown below instead of as in the patent:

column ll, lines 24, 36 and 49 for "alka-nol'fi read alkanoyl lines 28 to 35 the formula should appear as shown below instead of as in the patent:

each occurrence same column 11, lines 40 to 48, the formula should appear as shown below instead of as in the patent:

column 12, line 13, for "alaknol" read alkanoyl Signed and sealed this 17th day of September 1963.,

(SEAL) Attest:

ERNEST W. SWIDER DAVID L LADD Attesting Officer Commissioner of Patents 

3. THE COMPOUND 17B-HYDROXY-7A-METHYLTHIOANDROSTANO-(3,2-C)PYRAZOLE. 